Human PFKM Baculovirus-Insect cells Overexpression Lysate


Human PFKM Baculovirus-Insect cells Overexpression Lysate: 产品信息

This Human PFKM overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of PFKM protein (Cat: 14133-H20B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the human PFKM (P08237-1) (Thr2-Val780) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human PFKM /GST chimera consists of 1016 amino acids and has a calculated molecular mass of 112.9 kDa. The recombinant protein migrates as an approximately 113 kDa band in SDS-PAGE under reducing conditions.

Human PFKM Baculovirus-Insect cells Overexpression Lysate: 使用指南

Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human PFKM Baculovirus-Insect cells Overexpression Lysate: 别称

Human ATP-PFK Overexpression Lysate; Human GSD7 Overexpression Lysate; Human PFK-1 Overexpression Lysate; Human PFK1 Overexpression Lysate; Human PFKA Overexpression Lysate; Human PFKX Overexpression Lysate; Human PPP1R122 Overexpression Lysate

PFKM 背景信息

PFK1, also known as PFKM, is a regulatory glycolytic enzyme. PFK1 converts fructose 6-phosphate and ATP into fructose 1,6-bisphosphate (through PFK-1), fructose 2,6-bisphosphate (through PFK-2) and ADP. It is a muscle-type isozyme. There are three phosphofructokinase isozymes in humans: muscle, liver and platelet. These isozymes function as subunits of the mammalian tetramer phosphofructokinase, which catalyzes the phosphorylation of fructose-6-phosphate to fructose-1,6-bisphosphate. Mutations in PFK1 gene have been related to glycogen storage disease type VII, also identified as Tarui disease.
phosphofructokinase, muscle
  • Yamasaki T., et al.,(1991), Structure of the entire human muscle phosphofructokinase-encoding gene: a two-promoter system. Gene 104:277-282.
  • Sharma P.M., et al., (1989), Cloning and expression of a human muscle phosphofructokinase cDNA.Gene 77:177-183.
  • Nakajima H., et al.,(1987), Cloning of human muscle phosphofructokinase cDNA.FEBS Lett. 223:113-116.
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